Isolation and partial characterization of the lipoprotein families A and A-I from high-density lipoproteins of human serum.
Procedures for the isolation of two lipoprotein fractions from plasma high-density lipoproteins (HDL), characterized by apolipoprotein A-I and apolipoprotein A-I together with apolipoprotein A-II, have been elaborated. Apolipoprotein A-I was identified as the protein moiety of one of these fractions (lipoprotein A-I) with polyacrylamide gel electrophoresis (at basic and acidic pH, as well as in the presence of sodium dodecyl sulphate), immuno-double-diffusion, and amino acid analysis. Apolipoproteins A-I and A-II were identified as the protein moiety of the other fraction (lipoprotein A) with polyacrylamide gel electrophoresis (basic and acidic pH) and immuno-double-diffusion. Lipoprotein A-I consisted of spherical particles with a diameter similar to that of HDL as judged from negative strains in the transmission electron microscope. The diameter was estimated to be 8.7 nm from gel chromatography. Lipoprotein A-I migrated in the HDL position on crossed immunoelectrophoresis. On iso-electric focusing lipoprotein A-I appeared as multiple bands in the pH range 5.05-5.55. Lipoprotein A-I had the density of an HDL-2 fraction (rho: 1.063-1.105). Lipoprotein A consisted of spherical particles with a diameter similar to that of HDL, as judged from negative strains in the transmission electron microscope. The diameter was estimated to be 7.9 nm from gel chromatography. The molar ratio between the A-I and A-II polypeptides was estimated to 1.3:1 with electroimmunoassay and calculations from the amino acid compositions. Lipoprotein A migrated in the position of HDL on crossed immuno-electrophoresis. On iso-electric focusing lipoprotein A appeared as one major and two minor bands in the pH range 5.10-5.30. Lipoprotein A had the hydrated density of an HDL-2 fraction.
D000328:Adult; D016632:Apolipoprotein A-I; D001053:Apolipoproteins / Q000097:blood / Q000302:isolation & purification*; D001054:Apolipoproteins A; D001059:Apoproteins / Q000097:blood; D002499:Centrifugation, Density Gradient; D002784:Cholesterol / Q000032:analysis; D002846:Chromatography, Affinity / Q000379:methods; D006801:Humans; D005779:Immunodiffusion; D007123:Immunoelectrophoresis, Two-Dimensional; D008075:Lipoproteins, HDL / Q000097:blood*; D008297:Male; D008854:Microscopy, Electron; D008970:Molecular Weight; D010743:Phospholipids / Q000032:analysis; D014280:Triglycerides / Q000032:analysis
D016632:Apolipoprotein A-I; D001053:Apolipoproteins; D001054:Apolipoproteins A; D001059:Apoproteins; D008075:Lipoproteins, HDL; D010743:Phospholipids; D014280:Triglycerides; D002784:Cholesterol